Characterization of the bovine pregnancy-associated glycoprotein gene family – analysis of gene sequences, regulatory regions within the promoter and expression of selected genes

<p>Abstract</p> <p>Background</p> <p>The Pregnancy-associated glycoproteins (PAGs) belong to a large family of aspartic peptidases expressed exclusively in the placenta of species in the <it>Artiodactyla </it>order. In cattle, the <it>PAG </it>gene family is comprised of at least 22 transcribed genes, as well as some variants. Phylogenetic analyses have shown that the PAG family segregates into 'ancient' and 'modern' groupings. Along with sequence differences between family members, there are clear distinctions in their spatio-temporal distribution and in their relative level of expression. In this report, 1) we performed an <it>in silico </it>analysis of the bovine genome to further characterize the <it>PAG </it>gene family, 2) we scrutinized proximal promoter sequences of the <it>PAG </it>genes to evaluate the evolution pressures operating on them and to identify putative regulatory regions, 3) we determined relative transcript abundance of selected <it>PAGs </it>during pregnancy and, 4) we performed preliminary characterization of the putative regulatory elements for one of the candidate PAGs, <it>bovine </it>(<it>bo</it>) <it>PAG-2</it>.</p> <p>Results</p> <p>From our analysis of the bovine genome, we identified 18 distinct <it>PAG </it>genes and 14 pseudogenes. We observed that the first 500 base pairs upstream of the translational start site contained multiple regions that are conserved among all <it>boPAGs</it>. However, a preponderance of conserved regions, that harbor recognition sites for putative transcriptional factors (TFs), were found to be unique to the modern <it>boPAG </it>grouping, but not the ancient <it>boPAGs</it>. We gathered evidence by means of Q-PCR and screening of EST databases to show that <it>boPAG-2 </it>is the most abundant of all <it>boPAG </it>transcripts. Finally, we provided preliminary evidence for the role of ETS- and DDVL-related TFs in the regulation of the <it>boPAG-2 </it>gene.</p> <p>Conclusion</p> <p><it>PAGs </it>represent a relatively large gene family in the bovine genome. The proximal promoter regions of these genes display differences in putative TF binding sites, likely contributing to observed differences in spatial and temporal expression. We also discovered that <it>boPAG-2 </it>is the most abundant of all boPAG transcripts and provided evidence for the role of ETS and DDVL TFs in its regulation. These experiments mark the crucial first step in discerning the complex transcriptional regulation operating within the <it>boPAG </it>gene family.</p

Ethnic-Specific Differences in Vitamin D Status Is Associated with Adiposity

BackgroundLow circulating 25 hydroxyvitamin D [25(OH)D] concentrations are common in obesity (BMI ≥30 kg/m2) and a negative relationship with body fat distribution has recently been reported. Ethnic-specific differences in body fat distribution have been described with South Asians are reported to have greater visceral adipose tissue (VAT), which could influence circulating 25(OH)D concentrations. The objective of this study is to investigate the relationship between plasma 25(OH)D, adiposity, and body fat distribution in Europeans and South Asians.Methods/Principal Findings187 Europeans and 192 South Asians were assessed for demographics, anthropometrics, and plasma 25(OH)D concentrations. Subcutaneous adipose tissue (SAT) and VAT were quantified by CT scan, and percent body fat by DEXA. Data were assessed by general linear models. South Asians had lower (P&lt;0.001) plasma 25(OH)D concentrations and higher VAT (P = 0.04) than Europeans. Plasma 25(OH)D concentrations were negatively (P&lt;0.05) associated with BMI, waist circumference, percent body fat, total adipose tissue, VAT, and SAT in unadjusted models and negatively (P&lt;0.05) associated with VAT, SAT, and percent body fat after adjusting for BMI, ethnicity, age, and season of blood collection in males and females. When percent body fat, VAT, and SAT were included in the same model, only VAT remained negatively (P&lt;0.05) associated with plasma 25(OH)D concentrations. Ethnicity remained significant in all models (P&lt;0.001).ConclusionCompared to other adipose tissue compartments, VAT may have a distinct role in determining plasma 25(OH)D concentrations, which may account for the lower levels in South Asians

Early sex work initiation and condom use among alcohol-using female sex workers in Mombasa, Kenya: a cross-sectional analysis

Early initiation of sex work is prevalent among female sex workers (FSWs) worldwide. The objectives of this study were to investigate if early initiation of sex work was associated with: (1) consistent condom use, (2) condom negotiation self-efficacy or (3) condom use norms among alcohol-using FSWs in Mombasa, Kenya

Co-culture of Hepatocytes and Kupffer Cells as an In Vitro Model of Inflammation and Drug-Induced Hepatotoxicity

Immune-mediated drug-induced hepatotoxicity is often unrecognized as a potential mode of action due to the lack of appropriate in vitro models. We have established an in vitro rat donor-matched hepatocyte and Kupffer cell co-culture (HKCC) model to study immune-related responses to drug exposure. Optimal cell culture conditions were identified for the maintenance of co-cultures based on cell longevity, monolayer integrity, and cytokine response after lipopolysaccharide (LPS) exposure. Hepatocyte monocultures and HKCCs were then used to test a subset of compounds associated with hepatotoxic effects with or without LPS. Cytokine levels and metabolic activity (cytochrome P450 3A [Cyp3A]) were measured after a 48-h exposure to monitor endotoxin-induced changes in acute phase and functional end points. LPS-activated HKCCs, but not hepatocyte monocultures, treated with trovafloxacin or acetaminophen, compounds associated with immune-mediated hepatotoxicity, showed LPS-dependent decreases in interleukin-6 production with concomitant increases in Cyp3A activity. Differential endotoxinand model-dependent alterations were observed in cytokine profiles and Cyp3A activity levels that corresponded to specific compounds. These results indicate the utility of the HKCC model system to discern compound-specific effects that may lead to enhanced or mitigate hepatocellular injury due to innate or adaptive immune responses

Early Sex Work Initiation and Violence against Female Sex Workers in Mombasa, Kenya

Between 20 and 40 % of female sex workers (FSWs) began sex work before age 18. Little is known concerning whether early initiation of sex work impacts later experiences in adulthood, including violence victimization. This paper examines the relationship between early initiation of sex work and violence victimization during adulthood. The sample included 816 FSWs in Mombasa, Kenya, recruited from HIV prevention drop-in centers who were 18 years or older and moderate-risk drinkers. Early initiation was defined as beginning sex work at 17 or younger. Logistic regression modeled recent violence as a function of early initiation, adjusting for drop-in center, age, education, HIV status, supporting others, and childhood abuse. Twenty percent of the sample reported early initiation of sex work. Although both early initiators and other FSWs reported commonly experiencing recent violence, early initiators were significantly more likely to experience recent physical and sexual violence and verbal abuse from paying partners. Early initiation was not associated with physical or sexual violence from non-paying partners. Many FSWs begin sex work before age 18. Effective interventions focused on preventing this are needed. In addition, interventions are needed to prevent violence against all FSWs, in particular, those who initiated sex work during childhood or adolescence

Equine thermoregulatory responses during summertime road transport and stall confinement

Thermoregulatory responses of horses subjected to summer-time road transport and stall confinement were investigated in this study. Six mature geldings were transported 168 km in a 4-horse trailer and were monitored while tethered in their stalls, on alternate days. Core body temperature (GT) demonstrated negligible response during transport, but GT following transport was higher than GT for non-transport. GT tended to increase with increased temperature humidity index (THI). THI within the trailer was greatest for positions near the front, and was influenced by daily weather which varied over experiment days from heat stress conditions to moderate discomfort

Sex differences in the ability of corticostriatal oscillations to predict rodent alcohol consumption

Background: Although male and female rats differ in their patterns of alcohol use, little is known regarding the neural circuit activity that underlies these differences in behavior. The current study used a machine learning approach to characterize sex differences in local field potential (LFP) oscillations that may relate to sex differences in alcohol-drinking behavior. Methods: LFP oscillations were recorded from the nucleus accumbens shell and the rodent medial prefrontal cortex of adult male and female Sprague-Dawley rats. Recordings occurred before rats were exposed to alcohol (n = 10/sex × 2 recordings/rat) and during sessions of limited access to alcohol (n = 5/sex × 5 recordings/rat). Oscillations were also recorded from each female rat in each phase of estrous prior to alcohol exposure. Using machine learning, we built predictive models with oscillation data to classify rats based on: (1) biological sex, (2) phase of estrous, and (3) alcohol intake levels. We evaluated model performance from real data by comparing it to the performance of models built and tested on permutations of the data. Results: Our data demonstrate that corticostriatal oscillations were able to predict alcohol intake levels in males (p \u3c 0.01), but not in females (p = 0.45). The accuracies of models predicting biological sex and phase of estrous were related to fluctuations observed in alcohol drinking levels; females in diestrus drank more alcohol than males (p = 0.052), and the male vs. diestrus female model had the highest accuracy (71.01%) compared to chance estimates. Conversely, females in estrus drank very similar amounts of alcohol to males (p = 0.702), and the male vs. estrus female model had the lowest accuracy (56.14%) compared to chance estimates. Conclusions: The current data demonstrate that oscillations recorded from corticostriatal circuits contain significant information regarding alcohol drinking in males, but not alcohol drinking in females. Future work will focus on identifying where to record LFP oscillations in order to predict alcohol drinking in females, which may help elucidate sex-specific neural targets for future therapeutic development

The impact of an alcohol harm reduction intervention on interpersonal violence and engagement in sex work among female sex workers in Mombasa, Kenya: Results from a randomized controlled trial

To evaluate whether an alcohol harm reduction intervention was associated with reduced interpersonal violence or engagement in sex work among female sex workers (FSWs) in Mombasa, Kenya